Gene delivery systems are one of key issues that limit the development of gene therapy. The novel non-viral gene delivery systems fabricated via supramolecular assem- bly have begun to show increasing promising and ap...Gene delivery systems are one of key issues that limit the development of gene therapy. The novel non-viral gene delivery systems fabricated via supramolecular assem- bly have begun to show increasing promising and applica- tions in gene therapy due to its suitable nanometric size, controllable structure and excellent biocompatibility. In this review, the fundamental and recent progress of non-viral gene supramolecular assembly is reviewed. Artificial vi- ruses——the future direction of non-viral gene delivery sys- tems are also described.展开更多
RGD-containing peptide （ K16-GRGDSPC） , characterized as non-viral gene vectors, was fabricated to modify the surface of PLGA-[ASP- PEG] matrix, which offered the foundation for gene transfer with porous matrix of g...RGD-containing peptide （ K16-GRGDSPC） , characterized as non-viral gene vectors, was fabricated to modify the surface of PLGA-[ASP- PEG] matrix, which offered the foundation for gene transfer with porous matrix of gene activated later. Peptide was synthesized and matrix was executed into chips A, B and chip C. Chip C was regarded as control. Chips A and B were reacted with cross-linker. Then chip A was reacted with peptide. MS and HPLC were ased to detect the .14W and purity of peptide. Sulphur, existing on the surface of biomaterials, was detected by XPS. The purity of un-reacted peptide in residual solution was detected by a spectrophotometer. HPLC shows that the peptide purity was 94%- 95% , and MS shows that the MW was 2 741. 3307. XPS reveals that the binding energy of sulphur was 164 eV and the ratio of carbon to sulphur （C/S） was 99. 746 ：0. 1014 in reacted chip A. The binding energy of sulphur in reacted chip B was 164 eV and 162 eV, C/ S was 99.574：0.4255, aM there was no sulphur in chip C. Peptide was manufactured and linked to the surface of biomimetic and 3-D matrix, which offered the possibilities for gene transfer and tissue engineering with this new kind of non-viral gene vector.展开更多
Safe and effective gene therapy approaches require targeted tissue-specific transfer of a therapeutic transgene.Besides traditional approaches, such as transcriptional and transductional targeting, micro RNA-dependent...Safe and effective gene therapy approaches require targeted tissue-specific transfer of a therapeutic transgene.Besides traditional approaches, such as transcriptional and transductional targeting, micro RNA-dependent posttranscriptional suppression of transgene expression has been emerging as powerful new technology to increase the specificity of vector-mediated transgene expression. Micro RNAs are small non-coding RNAs and often expressed in a tissue-, lineage-, activation- or differentiation-specific pattern. They typically regulate gene expression by binding to imperfectly complementary sequences in the 3' untranslated region(UTR) of the m RNA. To control exogenous transgene expression, tandem repeats of artificial micro RNA target sites are usually incorporated into the 3' UTR of the transgene expression cassette, leading to subsequent degradation of transgene m RNA in cel s expressing the corresponding micro RNA. This targeting strategy, first shown for lentiviral vectors in antigen presenting cells, has now been used for tissue-specific expression of vector-encoded therapeutic transgenes, to reduce immune response against the transgene, to control virus tropism for oncolytic virotherapy, to increase safety of live attenuated virus vaccines and to identify and select cell subsets for pluripotent stem cell therapies, respectively. This review provides an introduction into the technical mechanism underlying micro RNA-regulation, highlights new developments in this field and gives an overview of applications of micro RNA-regulated viral vectors for cardiac, suicide gene cancer and hematopoietic stem cell therapy, as well as for treatment of neurological and eye diseases.展开更多
目的:基于网络药理学与GEO数据库探讨心瘅方治疗病毒性心肌炎的microRNA-mRNA调控机制。方法:使用中药系统药理学数据库与分析平台(traditional Chinese medicine systems pharmacology database and analysis platform,TCMSP)筛选心瘅...目的:基于网络药理学与GEO数据库探讨心瘅方治疗病毒性心肌炎的microRNA-mRNA调控机制。方法:使用中药系统药理学数据库与分析平台(traditional Chinese medicine systems pharmacology database and analysis platform,TCMSP)筛选心瘅方中药物活性成分及其靶基因;利用GEO数据库筛选病毒性心肌炎与健康人的差异microRNA;利用FunRich3.1.3对差异microRNA进行mRNA富集,并与药物靶基因取交集基因;通过蛋白-蛋白互作网络(protein-protein interactions,PPI)以及cytoNCA筛选出核心基因;运用R软件对核心基因进行基因本体(gene ontology,GO)和京都基因与基因组百科全书(kyoto encyclopedia of genes and genomes,KEGG)通路富集分析。结果:心瘅方中药物活性成分共199个,潜在作用靶点490个,GEO数据库Series GSE148153符合要求,通过筛选得到差异microRNA 438个,50个microRNA与核心基因有对应关系,上调基因45个,下调基因5个;筛选得到交集基因207个,其中33个为核心基因;GO功能注释主要富集在肌细胞增殖、MAP激酶活性等生物学过程,并且主要富集在PI3K/Akt、人巨细胞病毒感染、白细胞介素17等信号通路。结论:心瘅方调控microRNA-mRNA过程治疗病毒性心肌炎与中药成分干预肌细胞增殖、MAP激酶活性等生物学过程和PI3K/Akt、人巨细胞病毒感染、肿瘤坏死因子信号通路等有关,此过程可能涉及hsa-miR-15b-5p和hsa-miR-21-5p等外泌体的表达。展开更多
基金This work was fnancially supported by the Natu—ral Science Foundation of Zhejiang Province(Grant No.M503151)the National Natural Science Foundation of China(Grant No 50403021).
文摘Gene delivery systems are one of key issues that limit the development of gene therapy. The novel non-viral gene delivery systems fabricated via supramolecular assem- bly have begun to show increasing promising and applica- tions in gene therapy due to its suitable nanometric size, controllable structure and excellent biocompatibility. In this review, the fundamental and recent progress of non-viral gene supramolecular assembly is reviewed. Artificial vi- ruses——the future direction of non-viral gene delivery sys- tems are also described.
文摘RGD-containing peptide （ K16-GRGDSPC） , characterized as non-viral gene vectors, was fabricated to modify the surface of PLGA-[ASP- PEG] matrix, which offered the foundation for gene transfer with porous matrix of gene activated later. Peptide was synthesized and matrix was executed into chips A, B and chip C. Chip C was regarded as control. Chips A and B were reacted with cross-linker. Then chip A was reacted with peptide. MS and HPLC were ased to detect the .14W and purity of peptide. Sulphur, existing on the surface of biomaterials, was detected by XPS. The purity of un-reacted peptide in residual solution was detected by a spectrophotometer. HPLC shows that the peptide purity was 94%- 95% , and MS shows that the MW was 2 741. 3307. XPS reveals that the binding energy of sulphur was 164 eV and the ratio of carbon to sulphur （C/S） was 99. 746 ：0. 1014 in reacted chip A. The binding energy of sulphur in reacted chip B was 164 eV and 162 eV, C/ S was 99.574：0.4255, aM there was no sulphur in chip C. Peptide was manufactured and linked to the surface of biomimetic and 3-D matrix, which offered the possibilities for gene transfer and tissue engineering with this new kind of non-viral gene vector.
基金Supported by The Deutsche Forschungsgemeinschaft,Nos.FE785/2-2 and FE785/4-1the Bundesministerium für Bildung und Entwicklung,No.031A331
文摘Safe and effective gene therapy approaches require targeted tissue-specific transfer of a therapeutic transgene.Besides traditional approaches, such as transcriptional and transductional targeting, micro RNA-dependent posttranscriptional suppression of transgene expression has been emerging as powerful new technology to increase the specificity of vector-mediated transgene expression. Micro RNAs are small non-coding RNAs and often expressed in a tissue-, lineage-, activation- or differentiation-specific pattern. They typically regulate gene expression by binding to imperfectly complementary sequences in the 3' untranslated region(UTR) of the m RNA. To control exogenous transgene expression, tandem repeats of artificial micro RNA target sites are usually incorporated into the 3' UTR of the transgene expression cassette, leading to subsequent degradation of transgene m RNA in cel s expressing the corresponding micro RNA. This targeting strategy, first shown for lentiviral vectors in antigen presenting cells, has now been used for tissue-specific expression of vector-encoded therapeutic transgenes, to reduce immune response against the transgene, to control virus tropism for oncolytic virotherapy, to increase safety of live attenuated virus vaccines and to identify and select cell subsets for pluripotent stem cell therapies, respectively. This review provides an introduction into the technical mechanism underlying micro RNA-regulation, highlights new developments in this field and gives an overview of applications of micro RNA-regulated viral vectors for cardiac, suicide gene cancer and hematopoietic stem cell therapy, as well as for treatment of neurological and eye diseases.
文摘目的:基于网络药理学与GEO数据库探讨心瘅方治疗病毒性心肌炎的microRNA-mRNA调控机制。方法:使用中药系统药理学数据库与分析平台(traditional Chinese medicine systems pharmacology database and analysis platform,TCMSP)筛选心瘅方中药物活性成分及其靶基因;利用GEO数据库筛选病毒性心肌炎与健康人的差异microRNA;利用FunRich3.1.3对差异microRNA进行mRNA富集,并与药物靶基因取交集基因;通过蛋白-蛋白互作网络(protein-protein interactions,PPI)以及cytoNCA筛选出核心基因;运用R软件对核心基因进行基因本体(gene ontology,GO)和京都基因与基因组百科全书(kyoto encyclopedia of genes and genomes,KEGG)通路富集分析。结果:心瘅方中药物活性成分共199个,潜在作用靶点490个,GEO数据库Series GSE148153符合要求,通过筛选得到差异microRNA 438个,50个microRNA与核心基因有对应关系,上调基因45个,下调基因5个;筛选得到交集基因207个,其中33个为核心基因;GO功能注释主要富集在肌细胞增殖、MAP激酶活性等生物学过程,并且主要富集在PI3K/Akt、人巨细胞病毒感染、白细胞介素17等信号通路。结论:心瘅方调控microRNA-mRNA过程治疗病毒性心肌炎与中药成分干预肌细胞增殖、MAP激酶活性等生物学过程和PI3K/Akt、人巨细胞病毒感染、肿瘤坏死因子信号通路等有关,此过程可能涉及hsa-miR-15b-5p和hsa-miR-21-5p等外泌体的表达。
